NOTE: All solutions, equipment, and tissue culture materials coming into contact with cells must be sterile, and proper sterile
technique should be used accordingly. Solutions and materials should be free of endotoxin or other contaminants that may drive
DC maturation (take particular care in selecting protein preparations, e.g., bovine serum albumin or antigen preparations,
as these are frequently contaminated with endotoxin, which may introduce problems and artifacts).NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions, equipment, and tissue materials coming into contact with cells must be sterile, and proper sterile technique
should be used accordingly. Solutions and materials should be free of endotoxin or other contaminants that may drive DC maturation
(take particular care in selecting protein preparations, e.g., bovine serum albumin or antigen preparations, as these are
frequently contaminated with endotoxin, which may introduce problems and artifacts).NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions and tissue culture materials coming into contact with cells must be sterile, and proper sterile technique should
be used accordingly. Solutions and materials should be free of endotoxin or other contaminants that may drive DC maturation
(take particular care in selecting protein preparations, e.g., bovine serum albumin or antigen preparations, as these are
frequently contaminated with endotoxin, which may introduce problems and artifacts).

NOTE: All solutions and tissue culture materials coming into contact with cells must be sterile, and proper sterile technique should
be used accordingly.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

Support Protocol 2: Freezing PBMCs, Monocytes, or B Cells

Materials

PBMCs, monocytes, or B cells to be frozen (see appropriate protocols in this unit)

NOTE: All solutions and tissue culture materials coming into contact with cells must be sterile, and proper sterile technique should
be used accordingly. Solutions and materials should be free of endotoxin or other contaminants that may stimulate monocytes
(take particular care in selecting protein preparations, e.g., bovine serum albumin or antigen preparations, as these are
frequently contaminated with endotoxin, which may introduce problems and artifacts).

NOTE: All solutions, equipment and tissue culture ware coming into contact with cells must be sterile, and proper sterile technique
should be used accordingly.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions and tissue culture ware coming into contact with cells must be sterile, and proper sterile technique should
be used accordingly.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions and tissue culture ware coming into contact with cells must be sterile, and proper sterile technique should
be used accordingly.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions, equipment and tissue culture materials coming into contact with cells must be sterile, and proper sterile
technique should be used accordingly.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions, equipment and tissue culture materials coming into contact with cells must be sterile, and proper sterile
technique should be used accordingly. Listeria monocytogenes is a Biohazard Level 2 human pathogen, and appropriate precautions and decontamination should be performed. Pregnant women
and immunocompromised individuals should not work with L. monocytogenes.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions, equipment and tissue culture materials coming into contact with cells must be sterile, and proper sterile
technique should be used accordingly.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions and tissue culture ware coming into contact with cells must be sterile, and proper sterile technique should
be used accordingly.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions and tissue culture materials coming into contact with cells must be sterile, and proper sterile technique should
be used accordingly.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.

NOTE: All solutions, equipment and tissue culture materials coming into contact with cells must be sterile, and proper sterile
technique should be used accordingly.NOTE: All culture incubations are performed in a humidified 37°C, 5% CO2 incubator unless otherwise specified.